Development and evaluation of a novel advanced lipoprotein test based on 2d diffusion orderen 1h nmr spectroscopy

La determinació de la mida i el nombre de lipoproteïnes utilitzant tests avançats de lipoproteïnes és d'un gran interès clínic ja que el nombre de partícules LDL s'ha posicionat com a millor predictor de risc cardiovascular que el colesterol LDL. Tanmateix, els tests avançats de lipoproteïnes actuals encara no s'han introduït en l'àmbit clínic en gran part per la falta d'una estandarització. En aquesta tesi presentem el test LipoScale, un nou test avançat de lipoproteïnes basat en espectroscopia de RMN de difusió 2D. Amb aquest test es pretén obtenir una millor caracterització de les lipoproteïnes plasmàtiques, tant el seu contingut lipídic com la seva mida i nombre de partícules, de manera que amb ell s'aconsegueixi una millor predicció del risc cardiovascular. Durant el desenvolupament del test s’han estudiat diferents patologies i cohorts dins del marc de les malalties metabòliques (les quals són un factor de risc de les malalties cardiovasculars). Entre les malalties estudiades destaquem la diabetis, la dislipèmia aterògena i la síndrome de l’ovari poliquístic (PCOS). A més, també s’han monitoritzat canvis en el perfil de les lipoproteïnes deguts a intervencions nutricionals i a l’exercici. La principal diferència entre la nostra aproximació i la dels mètodes actuals és que aquests últims utilitzen mètodes de RMN 1D estàndards, mentre que el nostre test està basat en l'ús de gradients de camps magnètic, els quals generen espectres 2D amb els que es pot obtenir informació directa i objectiva de la mida de les partícules lipoproteiques. Aquesta tesi ha generat diferents publicacions científiques així com també s'ha fet la sol•licitud d'una patent europea i s'ha creat una spin-off per comercialitzar el test.La determinación del tamaño y el número de lipoproteínas utilizando tests avanzados de lipoproteínas es de un gran interés clínico ya que el número de partículas LDL se ha posicionado como mejor predictor de riesgo cardiovascular que el colesterol LDL. Sin embargo, los tests avanzados de lipoproteínas actuales aún no se han introducido en el ámbito clínico en gran parte por la falta de una estandarización. En esta tesis presentamos el test LipoScale, un nuevo test avanzado de lipoproteínas basado en espectroscopía de RMN de difusión 2D. Con este test se pretende obtener una mejor caracterización de las lipoproteínas plasmáticas, tanto su contenido lipídico como su tamaño y número de partículas, por lo que con él se consiga una mejor predicción del riesgo cardiovascular. Durante el desarrollo del test se han estudiado diferentes patologías y cohortes dentro del marco de las enfermedades metabólicas (las cuales son un factor de riesgo de las enfermedades cardiovasculares). Entre las enfermedades estudiadas destacamos la diabetes, la dislipemia aterògena y el síndrome del ovario poliquístico (PCOS). Además, también se han monitorizado cambios en el perfil de las lipoproteínas debidos a intervenciones nutricionales y el ejercicio. La principal diferencia entre nuestra aproximación y la de los métodos actuales es que estos últimos utilizan métodos de RMN 1D estándar, mientras que nuestro test está basado en el uso de gradientes de campo magnético, los cuales generan espectros 2D con los que se puede obtener información directa y objetiva del tamaño de las partículas lipoproteicas. Esta tesis a generado diferentes publicaciones científicas así como también se ha hecho la solicitud de una patente europea y se ha creado una spin-off para comercializar el test.Determination of lipoprotein particle size and particle number using advanced lipoprotein analyses is of particular interest since the LDL particle number has been shown to improve cardiovascular disease risk prediction. Advanced lipoprotein tests (ALT), however, are not yet routinely introduced in clinical practice partly due to the lack of standardization. This thesis presents the LipoScale test, a novel advanced lipoprotein test based on 2D diffusion-ordered 1H NMR spectroscopy. This test is to obtain a better characterization of plasma lipoproteins in terms of their lipid content, particle size and particle number that will allow a better assessment of cardiovascular risk. During the development of the test various diseases and cohorts were studied in the context of metabolic diseases (which are a risk factor for cardiovascular disease). Among the diseases studied we highlight diabetes, atherogenic dyslipidemia and polycystic ovary syndrome (PCOS). In addition, changes were also monitored in the lipoprotein profile due to nutritional interventions and exercise. The main difference between our approach and the current NMR methods is that the latter use standard 1D methods, whereas our test is based on the use of magnetic field gradients, which generate the 2D spectra that can be used to get direct and objective information on lipoprotein particle sizes. This thesis generated various scientific publications, includes an application for a European patent and a spin-off has been created to commercialize the test

The Neanderthal Meal: A New Perspective Using Faecal Biomarkers

Neanderthal dietary reconstructions have, to date, been based on indirect evidence and may underestimate the significance of plants as a food source. While zooarchaeological and stable isotope data have conveyed an image of Neanderthals as largely carnivorous, studies on dental calculus and scattered palaeobotanical evidence suggest some degree of contribution of plants to their diet. However, both views remain plausible and there is no categorical indication of an omnivorous diet. Here we present direct evidence of Neanderthal diet using faecal biomarkers, a valuable analytical tool for identifying dietary provenance. Our gas chromatography-mass spectrometry results from El Salt (Spain), a Middle Palaeolithic site dating to ca. 50,000 yr. BP, represents the oldest positive identification of human faecal matter. We show that Neanderthals, like anatomically modern humans, have a high rate of conversion of cholesterol to coprostanol related to the presence of required bacteria in their guts. Analysis of five sediment samples from different occupation floors suggests that Neanderthals predominantly consumed meat, as indicated by high coprostanol proportions, but also had significant plant intake, as shown by the presence of 5β-stigmastanol. This study highlights the applicability of the biomarker approach in Pleistocene contexts as a provider of direct palaeodietary information and supports the opportunity for further research into cholesterol metabolism throughout human evolution.NASA Astrobiology Institute (Grant NNA13AA90A

Lipoprotein hydrophobic core lipids are partially extruded to surface in smaller HDL : "Herniated" HDL, a common feature in diabetes

Recent studies have shown that pharmacological increases in HDL cholesterol concentrations do not necessarily translate into clinical benefits for patients, raising concerns about its predictive value for cardiovascular events. Here we hypothesize that the size-modulated lipid distribution within HDL particles is compromised in metabolic disorders that have abnormal HDL particle sizes, such as type 2 diabetes mellitus (DM2). By using NMR spectroscopy combined with a biochemical volumetric model we determined the size and spatial lipid distribution of HDL subclasses in a cohort of 26 controls and 29 DM2 patients before and after two drug treatments, one with niacin plus laropiprant and another with fenofibrate as an add-on to simvastatin. We further characterized the HDL surface properties using atomic force microscopy and fluorescent probes to show an abnormal lipid distribution within smaller HDL particles, a subclass particularly enriched in the DM2 patients. The reduction in the size, force cholesterol esters and triglycerides to emerge from the HDL core to the surface, making the outer surface of HDL more hydrophobic. Interestingly, pharmacological interventions had no effect on this undesired configuration, which may explain the lack of clinical benefits in DM2 subjects

Metabolomics Reveals Reduction of Metabolic Oxidation in Women with Polycystic Ovary Syndrome after Pioglitazone-Flutamide-Metformin Polytherapy

Polycystic ovary syndrome (PCOS) is a variable disorder characterized by a broad spectrum of anomalies, including hyperandrogenemia, insulin resistance, dyslipidemia, body adiposity, low-grade inflammation and increased cardiovascular disease risks. Recently, a new polytherapy consisting of low-dose flutamide, metformin and pioglitazone in combination with an estro-progestagen resulted in the regulation of endocrine clinical markers in young and non-obese PCOS women. However, the metabolic processes involved in this phenotypic amelioration remain unidentified. In this work, we used NMR and MS-based untargeted metabolomics to study serum samples of young non-obese PCOS women prior to and at the end of a 30 months polytherapy receiving low-dose flutamide, metformin and pioglitazone in combination with an estro-progestagen. Our results reveal that the treatment decreased the levels of oxidized LDL particles in serum, as well as downstream metabolic oxidation products of LDL particles such as 9- and 13-HODE, azelaic acid and glutaric acid. In contrast, the radiuses of small dense LDL and large HDL particles were substantially increased after the treatment. Clinical and endocrine-metabolic markers were also monitored, showing that the level of HDL cholesterol was increased after the treatment, whereas the level of androgens and the carotid intima-media thickness were reduced. Significantly, the abundance of azelaic acid and the carotid intima-media thickness resulted in a high degree of correlation. Altogether, our results reveal that this new polytherapy markedly reverts the oxidant status of untreated PCOS women, and potentially improves the pro-atherosclerosis condition in these patients

Metabolomics reveals impaired maturation of HDL particles in adolescents with hyperinsulinaemic androgen excess.

Hyperinsulinaemic androgen excess (HIAE) in prepubertal and pubertal girls usually precedes a broader pathological phenotype in adulthood that is associated with anovulatory infertility, metabolic syndrome and type 2 diabetes. The metabolic derangements that determine these long-term health risks remain to be clarified. Here we use NMR and MS-based metabolomics to show that serum levels of methionine sulfoxide in HIAE girls are an indicator of the degree of oxidation of methionine-148 residue in apolipoprotein-A1. Oxidation of apo-A1 in methionine-148, in turn, leads to an impaired maturation of high-density lipoproteins (HDL) that is reflected in a decline of large HDL particles. Notably, such metabolic alterations occur in the absence of impaired glucose tolerance, hyperglycemia and hypertriglyceridemia, and were partially restored after 18 months of treatment with a low-dose combination of pioglitazone, metformin and flutamide

A baseline metabolomic signature is associated with immunological CD4+ T-Cell recovery after 36 months of art in HIV-infected patients

Poor immunological recovery in treated HIV-infected patients is associated with greater morbidity and mortality. To date, predictive biomarkers of this incomplete immune reconstitution have not been established. We aimed to identify a baseline metabolomic signature associated with a poor immunological recovery after ART in order to envisage the underlying mechanistic pathways that influence the treatment response.Peer reviewe

A baseline metabolomic signature is associated with immunological CD4 + T-cell recovery after 36 months of antiretroviral therapy in HIV-infected patients

Poor immunological recovery in treated HIV-infected patients is associated with greater morbidity and mortality. To date, predictive biomarkers of this incomplete immune reconstitution have not been established. We aimed to identify a baseline metabolomic signature associated with a poor immunological recovery after antiretroviral therapy (ART) to envisage the underlying mechanistic pathways that influence the treatment response. This was a multicentre, prospective cohort study in ART-naive and a pre-ART low nadir (<200 cells/μl) HIV-infected patients (n = 64). We obtained clinical data and metabolomic profiles for each individual, in which low molecular weight metabolites, lipids and lipoproteins (including particle concentrations and sizes) were measured by NMR spectroscopy. Immunological recovery was defined as reaching CD4 + T-cell count at least 250 cells/μl after 36 months of virologically successful ART. We used univariate comparisons, Random Forest test and receiver-operating characteristic curves to identify and evaluate the predictive factors of immunological recovery after treatment. HIV-infected patients with a baseline metabolic pattern characterized by high levels of large high density lipoprotein (HDL) particles, HDL cholesterol and larger sizes of low density lipoprotein particles had a better immunological recovery after treatment. Conversely, patients with high ratios of non-HDL lipoprotein particles did not experience this full recovery. Medium very-low-density lipoprotein particles and glucose increased the classification power of the multivariate model despite not showing any significant differences between the two groups. In HIV-infected patients, a baseline healthier metabolomic profile is related to a better response to ART where the lipoprotein profile, mainly large HDL particles, may play a key role

RICORS2040 : The need for collaborative research in chronic kidney disease

Chronic kidney disease (CKD) is a silent and poorly known killer. The current concept of CKD is relatively young and uptake by the public, physicians and health authorities is not widespread. Physicians still confuse CKD with chronic kidney insufficiency or failure. For the wider public and health authorities, CKD evokes kidney replacement therapy (KRT). In Spain, the prevalence of KRT is 0.13%. Thus health authorities may consider CKD a non-issue: very few persons eventually need KRT and, for those in whom kidneys fail, the problem is 'solved' by dialysis or kidney transplantation. However, KRT is the tip of the iceberg in the burden of CKD. The main burden of CKD is accelerated ageing and premature death. The cut-off points for kidney function and kidney damage indexes that define CKD also mark an increased risk for all-cause premature death. CKD is the most prevalent risk factor for lethal coronavirus disease 2019 (COVID-19) and the factor that most increases the risk of death in COVID-19, after old age. Men and women undergoing KRT still have an annual mortality that is 10- to 100-fold higher than similar-age peers, and life expectancy is shortened by ~40 years for young persons on dialysis and by 15 years for young persons with a functioning kidney graft. CKD is expected to become the fifth greatest global cause of death by 2040 and the second greatest cause of death in Spain before the end of the century, a time when one in four Spaniards will have CKD. However, by 2022, CKD will become the only top-15 global predicted cause of death that is not supported by a dedicated well-funded Centres for Biomedical Research (CIBER) network structure in Spain. Realizing the underestimation of the CKD burden of disease by health authorities, the Decade of the Kidney initiative for 2020-2030 was launched by the American Association of Kidney Patients and the European Kidney Health Alliance. Leading Spanish kidney researchers grouped in the kidney collaborative research network Red de Investigación Renal have now applied for the Redes de Investigación Cooperativa Orientadas a Resultados en Salud (RICORS) call for collaborative research in Spain with the support of the Spanish Society of Nephrology, Federación Nacional de Asociaciones para la Lucha Contra las Enfermedades del Riñón and ONT: RICORS2040 aims to prevent the dire predictions for the global 2040 burden of CKD from becoming true

Sons al balcó: Soundscape Map of the Confinement in Catalonia

In this project, we aim to study the effect that the lockdown due to the COVID-19 pandemic has caused on the perception of noise in Catalonia. In Sons al Balc&oacute;, the research activities cohabit with the dynamic collaboration with citizens and other stakeholders to create social and environmental impact, to widen awareness and design tools to improve citizenship development and empowerment. The initial scientific hypothesis is that the annoyance coming from outdoor noise, minimized by the lockdown effect, could be associated with better perception of the soundscape by people. Sons al Balc&oacute; allows validating this hypothesis in two different ways. On the one hand, by means of subjective questionnaires conducted to people living in pre-defined diverse acoustic areas (urban, suburban and rural environments), and on the other hand, by the use of objective measurements of the noise levels, and the study of the soundscape in these areas, using short pieces of video collected by citizens. For this purpose, we designed an on line test to be conducted by any citizen aiming to contribute to this wide study for all the territory of Catalonia, both from rural areas and from cities. A communication campaign was conducted to reach a significant participation. During the lockdown, more than 350 questionnaires and videos were collected, and a first map of the soundscape of the confinement in Catalonia was depicted